Patent DE-102010044562-A1 (DE)
Title: Automated techniques of in vitro propagation of useful and ornamental plants based on somatic embryogenesis and adventitious process for forming homogenous plant-cluster, based on regeneration technique using stem cells of apical meristem
IPC: C12N5/04
Filed: 2010-12-08
Abstract: Automated techniques of in vitro propagation of useful and ornamental plants based on somatic embryogenesis and adventitious processes for forming homogenous plant-cluster, based on a regeneration technique using stem cells of apical meristem, whose directed proliferation under targeted expression of phytohormones leads to formation of homogeneous secondary embryoids with unlimited proliferation ability and further a nutrient medium is subjected, whose composition is adjusted such that an induction of defined differentiation is formed leading to a controllable plant regeneration, is claimed. Automated techniques of in vitro propagation of useful and ornamental plants on the basis of somatic embryogenesis and adventitious processes for forming homogenous plant-cluster, prefaced by organogenesis, based on a regeneration technique using stem cells of the apical meristem, whose directed proliferation under the targeted expression of phytohormones leads to the formation of homogeneous secondary embryoids with the property of unlimited ability to proliferate and further a nutrient medium is subjected, those composition is composed in such a way that an induction of defined differentiation is formed, which leads to a controllable plant regeneration, comprises: (i) isolating cell groups from the apical area of the meristem occupied with cells having the property of pluripotency according to the Figure 1 and 2 of the specification; isolating and explanting cells groups from the apical area of the meristem; placing the explanted cells in a nutrient medium N1, which is given in a sterilizable vascular growth system G1; operating an artificial plant species specific controlled transfer of the nutrient medium N1 with cytokinins, on the basis of a nutrient medium N1.1 with controllable cytokinin concentration; producing a direct somatic embryogenesis from the explanted stem cells of pluripotency-sector of homogeneous embryoids; automatically quantizing/portioning the embryoid miniature plant; automatically transplanting the quantized and segmented hypocotyl explants of developed embryoids miniature plants on the basis of a culture medium N2, which is given in a sterilizable vascular growth G2 with segmented implantation space; operating an artificial plant species specific controlled transfer of the nutrient medium N2 with auxins/cytokinins, on the basis of the nutrient medium N2.1 with controllable auxin-/cytokinin-concentration ratio; originating a generation process of the secondary somatic embryos in the form of a micro-plant cluster with self-sufficient individual plants; recording and evaluating the physiological growth and plant status of the micro-plant cluster with self-sufficient individual plants in a sensory on line manner; recording/localizing the micro-plant-cluster without ex vitro suitability in a sensory manner and removing the further growth sequence; and transplanting and mycorrhization of the ex vitro-suitable, in vitro developed plant-clusters with self-sufficient individual plants automated in the ex vitro environment of the natural habitat and/or adding with effective regenerative microorganisms (EMS).